Changes of content and mRNA expression of gelsolin and proliferation activity of T-lymphocyte in spleen of mice with severe burn injury
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摘要: 目的 研究重度烧伤小鼠脾脏凝溶胶蛋白(GSN)含量和mRNA表达及T淋巴细胞增殖活性的变化,选择GSN的最佳干预时间。 方法 取80只雄性BALB/c小鼠,按随机数字表法分为假伤组和烧伤组,每组40只。烧伤组小鼠造成背部15%TBSAⅢ度烫伤(下称烧伤),伤后即刻皮下注射生理盐水1 mL,背部涂适量碘伏防治感染,每日1次。假伤组小鼠致假伤,伤后不补液、不外涂碘伏。分别于伤后0(即刻)、8、24、48、72 h,每组各取8只小鼠,无菌留取脾脏,噻唑蓝比色法检测脾脏T淋巴细胞增殖活性,ELISA法检测脾脏组织中GSN含量,实时荧光定量RT-PCR法检测脾脏组织中GSN mRNA表达。对数据行析因设计方差分析、单因素方差分析、LSD检验及Bonferroni校正。 结果 (1)2组小鼠伤后0 h的脾脏T淋巴细胞增殖活性无明显差异(
P >0.05),假伤组小鼠伤后8、24、48、72 h脾脏T淋巴细胞增殖活性显著高于烧伤组(P 值均小于0.05)。假伤组小鼠伤后各时相点脾脏T淋巴细胞增殖活性无明显差异(F =0.756,P >0.05);烧伤组小鼠伤后8 h脾脏T淋巴细胞增殖活性为0.12±0.04,显著低于组内伤后0、24、48、72 h的0.73±0.07、0.56±0.07、0.51±0.09、0.59±0.07(P 值均小于0.05)。(2)2组小鼠伤后0 h脾脏组织中GSN含量无明显差异(P >0.05),假伤组小鼠伤后8、24、48、72 h脾脏组织中GSN含量显著高于烧伤组(P 值均小于0.05)。假伤组小鼠伤后各时相点脾脏组织中GSN含量无明显差异(F =1.083,P >0.05);烧伤组小鼠伤后8 h脾脏组织中GSN含量为(11.9±2.6)pg/mg,显著低于组内伤后0、24、48、72 h的(37.7±2.9)、(19.9±4.0)、(24.1±4.1)、(24.6±4.0)pg/mg(P 值均小于0.05)。(3)2组小鼠伤后0 h脾脏组织中GSN mRNA表达量无明显差异(P >0.05),假伤组小鼠伤后8、24、48、72 h脾脏组织中GSN mRNA表达量显著高于烧伤组(P 值均小于0.05)。假伤组小鼠伤后各时相点脾脏组织中GSN mRNA表达量无明显差异(F =0.413,P >0.05);烧伤组小鼠伤后8 h脾脏组织中GSN mRNA表达量为0.307±0.064,显著低于组内伤后0、24、48、72 h的0.944±0.023、0.625±0.091、0.744±0.104、0.821±0.072(P 值均小于0.05)。 结论 重度烧伤可导致小鼠脾脏GSN含量及mRNA表达、T淋巴细胞增殖活性显著下降,且三者均是在伤后8 h降至最低,可选择伤后8 h之前作为重度烧伤后GSN的最佳干预时间。Abstract: Objective To investigate the changes of content and mRNA expression of gelsolin and proliferation activity of T-lymphocyte in spleen of mice with severe burn injury, so as to determine the optimum intervention time of gelsolin. Methods Eighty male BALB/c mice were divided into sham injury group and burn group according to the random number table, with 40 mice in each group. Mice in burn group were inflicted with 15% total body surface area full-thickness scald (hereinafter referred to as burn) on the back. Immediately after injury, mice in burn group were hypodermic injected with 1 mL normal saline, with iodophor smeared on back once a day to prevent infection. Mice in sham injury group were sham injured without fluid infusion and smearing iodophor. At post injury hour (PIH) 0 (immediately), 8, 24, 48, and 72, spleen of 8 mice of each group were harvested aseptically, respectively. Proliferation activity of T-lymphocyte was determined with methyl-thiazolyl-tetrazolium colorimetry method; gelsolin content of spleen was determined with enzyme-linked immunosorbent assay; mRNA expression of gelsolin of spleen was determined with real-time fluorescent quantitative reverse transcription polymerase chain reaction. Data were processed with analysis of variance of factorial design, one-way analysis of variance, LSD test and Bonferroni correction. Results (1) There was no significant difference in proliferation activity of T-lymphocyte in spleen of mice in two groups at PIH 0 (P >0.05). Proliferation activity of T-lymphocyte in spleen of mice in sham injury group was significantly higher than that in burn group at PIH 8, 24, 48, and 72 (withP values below 0.05). There was no significant difference in proliferation activity of T-lymphocyte in spleen of mice in sham injury group at each time point post injury (F =0.756,P >0.05). Proliferation activity of T-lymphocyte in spleen of mice in burn group at PIH 8 was 0.12±0.04, significantly lower than that at PIH 0, 24, 48, and 72 in the same group (0.73±0.07, 0.56±0.07, 0.51±0.09, and 0.59±0.07, respectively, withP values below 0.05). (2) There was no significant difference in gelsolin content of spleen of mice in two groups at PIH 0 (P >0.05). Gelsolin content of spleen of mice in sham injury group was significantly higher than that in burn group at PIH 8, 24, 48, and 72 (withP values below 0.05). There was no significant difference in gelsolin content of spleen of mice in sham injury group at each time point post injury (F =1.083,P >0.05). Gelsolin content of spleen of mice in burn group at PIH 8 was (11.9±2.6) pg/mg, significantly lower than that at PIH 0, 24, 48, and 72 in the same group [(37.7±2.9), (19.9±4.0), (24.1±4.1), and (24.6±4.0) pg/mg, respectively, withP values below 0.05]. (3) There was no significant difference in mRNA expression of gelsolin of spleen of mice in two groups at PIH 0 (P >0.05). The mRNA expressions of gelsolin of spleen of mice in sham injury group were significantly higher than those in burn group at PIH 8, 24, 48, and 72 (withP values below 0.05). There was no significant difference in mRNA expression of gelsolin of spleen of mice in sham injury group at each time point post injury (F =0.413,P >0.05). The mRNA expression of gelsolin of spleen of mice in burn group at PIH 8 was 0.307±0.064, significantly lower than that at PIH 0, 24, 48, and 72 in the same group (0.944±0.023, 0.625±0.091, 0.744±0.104, and 0.821±0.072, respectively, withP values below 0.05). Conclusions Severe burn injury could induce decrease of proliferation activity of T-lymphocyte and content and mRNA expressions of gelsolin in spleen of mice, and all of them decreased into the lowest at PIH 8. Optimum intervention time of gelsolin for severe burn would be before PIH 8.-
Key words:
- Burns /
- Spleen /
- T-lymphocytes /
- Cell proliferation /
- Gene expression /
- Gelsolin
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